{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T19:00:27Z","timestamp":1785438027478,"version":"3.56.0"},"reference-count":34,"publisher":"MDPI AG","issue":"14","license":[{"start":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T00:00:00Z","timestamp":1784160000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"key project from the Sichuan Province of China","award":["2020YFH0020"],"award-info":[{"award-number":["2020YFH0020"]}]},{"name":"key project from the Sichuan Province of China","award":["2021YFYZ0030"],"award-info":[{"award-number":["2021YFYZ0030"]}]},{"name":"Chengdu International Technology Innovation Cooperation Program","award":["2025-GH02-00035-HZ"],"award-info":[{"award-number":["2025-GH02-00035-HZ"]}]},{"name":"key project of Sichuan University, sanyoukang Corp","award":["23H1442"],"award-info":[{"award-number":["23H1442"]}]},{"award":["23H1442"],"award-info":[{"award-number":["23H1442"]}],"id":[{"id":"https:\/\/ror.org\/011ashp19","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biology"],"abstract":"<jats:p>Porcine circovirus type 2 (PCV2) and Mycoplasma hyopneumoniae (Mhp) are major contributors to the porcine respiratory disease complex. Although PCV2-Mhp bivalent inactivated vaccines are useful for simultaneous disease control, their immunogenicity may be improved by adjuvant optimization. This study evaluated a composite adjuvant consisting of porcine interleukin-2 (IL-2), IL-4, IL-6 and MnJ(beta), a colloidal manganese adjuvant, in an initial murine immunogenicity model. Thirty female Kunming mice were assigned to three groups (n = 10\/group): bivalent antigen plus IL-2\/IL-4\/IL-6\/MnJ(beta), bivalent antigen plus MnJ(beta), or phosphate-buffered saline. Body weight, complete blood count, peripheral blood T- and B-cell subsets, PCV2-specific IgG and Mhp-specific indirect hemagglutination titers were monitored after primary and booster immunization. The composite formulation did not suppress body-weight gain or induce sustained abnormalities in erythrocyte- or platelet-related indices. WBC, neutrophil, lymphocyte and monocyte counts were elevated in group A at days 7 and 28 post-primary immunization, indicating transient immune activation. Day-56 flow cytometry indicated increased CD19+IgM-IgD- B-cell and effector\/memory T-cell-associated responses. PCV2-specific IgG increased from day 14 onward. At day 56, the OD450 value in group A reached 1.532 \u00b1 0.006, compared with 1.095 \u00b1 0.004 in group C1 and 0.102 \u00b1 0.002 in group C2, corresponding to approximately 1.40-fold and 15.09-fold higher levels than the MnJ(beta)-adjuvanted and PBS control groups, respectively. Mhp-specific IHA titers were also maintained at high levels after booster immunization; at day 56, group A showed a log2 endpoint titer of 13.00 \u00b1 0.00, corresponding to a GMT of 1:8192, whereas group C1 showed a log2 endpoint titer of 12.00 \u00b1 0.00, corresponding to a GMT of 1:4096, and group C2 remained negative. These results indicate that the IL-2\/IL-4\/IL-6\/MnJ(beta) composite adjuvant demonstrates potential for improving antibody and peripheral lymphocyte responses to PCV2-Mhp bivalent antigen, but protective efficacy must be confirmed in target-species challenge studies.<\/jats:p>","DOI":"10.3390\/biology15141163","type":"journal-article","created":{"date-parts":[[2026,7,16]],"date-time":"2026-07-16T08:08:34Z","timestamp":1784189314000},"page":"1163","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Porcine Interleukin-2, IL-4 and IL-6 Combined with a Colloidal Manganese Adjuvant Enhance PCV2-Mhp Bivalent Inactivated Vaccine Immunogenicity in Mice"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7303-2562","authenticated-orcid":false,"given":"Junjie","family":"Peng","sequence":"first","affiliation":[{"name":"National Engineering Research Center for Biomaterial, Sichuan University, Chengdu 610065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5172-2247","authenticated-orcid":false,"given":"Linhan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, China"},{"name":"Sichuan Sanyoukang Biotechnological Co., Ltd., Chengdu 610093, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dafang","family":"He","sequence":"additional","affiliation":[{"name":"Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, China"},{"name":"Sichuan Sanyoukang Biotechnological Co., Ltd., Chengdu 610093, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3296-3015","authenticated-orcid":false,"given":"Gang","family":"Wang","sequence":"additional","affiliation":[{"name":"National Engineering Research Center for Biomaterial, Sichuan University, Chengdu 610065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianglin","family":"Li","sequence":"additional","affiliation":[{"name":"Sichuan Academy of Animal Science, Chengdu 610066, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shanshan","family":"Zhu","sequence":"additional","affiliation":[{"name":"Sichuan University Library, Sichuan University, Chengdu 610065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-7954-4910","authenticated-orcid":false,"given":"Rong","family":"Gao","sequence":"additional","affiliation":[{"name":"Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,7,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.virusres.2011.10.007","article-title":"Porcine Circovirus Type 2 (PCV2) Infections: Clinical Signs, Pathology and Laboratory Diagnosis","volume":"164","year":"2012","journal-title":"Virus Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1146\/annurev-animal-031412-103720","article-title":"Porcine Circovirus Type 2 (PCV2): Pathogenesis and Interaction with the Immune System","volume":"1","author":"Meng","year":"2013","journal-title":"Annu. 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